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细胞功能测定

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Laboratory Diagnosis of Structural Hemoglobinopathies and Thalassemias by Capillary Isoelectric Focusing

Structural hemoglobinopathies and thalassemias are congenital hemoglobin (Hb) disorders that cause anemia, morbidity, and mortality resulting from abnormal Hb function. Structurally different normal Hb variants include HbA2, HbF (fetal hemoglobin), and HbA (adult hemog ...

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Steroid Analysis by Micellar Electrokinetic Capillary Chromatography

From the clinical perspective, steroids have always held a great deal of interest, since they are highly specific in their function. They do not have a general or systemic effect, but instead regulate specific physiological functions, such as sex differentiation, fetus implantation and gr ...

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Confirmation of the Presence of Drugs of Abuse in Urine

Urine screenings for drugs of abuse are performed for many reasons, two of the most common being workplace/employment screening and monitoring in drug-dependence treatment centers. In both, the initial screen is commonly an immunoassay. Negative results require no further action. Po ...

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Acetonitrile Stacking: Serum Phenobarbital as an Example

Capillary electrophoresis (CE) has several advantages, such as high resolution, speed, and low cost of operation. However, it suffers from two major drawbacks: poor detection limits and matrix effects. Several approaches have been used to overcome these two problems. Here, acetonitrile ...

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Serum Lamotrigine Analysis

Antiepileptic drugs vary greatly in chemical structure. Analysis of these compounds by capillary electrophoresis (CE) requires different conditions for each one, or for each group. Acidic drugs can be analyzed easily by capillary zone electrophoresis in borate buffers (1); the neutr ...

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Identification of Differentially Expressed Genes in Sorted Cell Populations by Two-Dimensional Gene-Expression Fingerprinting

Differential activity of genes is one of the major mechanisms underlying a vast array of biological phenomena. Classical genetic approaches (from phenotypes to genes) have proven their exquisite potential for dissection of complex signaling pathways regulating the development ...

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The Purification of Mouse Hematopoietic Stem Cells at Sequential Stages of Maturation

Hematopoietic stem cells (HSCs) are rare, self-renewing progenitors that give rise to all lineages of blood cells. HSCs can be found in all hematopoietic organs, from the para-aortic mesoderm (1),(2) and yolk sac (3),(4) in fetuses to the bone marrow (reviewed in ref. 5), blood and spleens of adults.

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Flow Cytometry and Immunoselection of Human Stem Cells

Human hematopoietic stem cells (HSCs) can be obtained from a variety of hematopoietic tissues, including bone marrow, blood, cord blood, and fetal liver. Various techniques have been used to fractionate hematopoietic cell populations based on differences in size and density, express ...

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Noninvasive Measurement of Hematopoietic Stem Cell Cycling and Turnover by In Vivo Bromodeoxyuridine Incorporation

The hematopoietic system comprises a concatenated series of stem- and transit-progenitor-cell compartments of progressively restricted potentiality and proliferative capacity (1–5). Analysis of hematopoietic regulation in transplantation models and in marrow rege ...

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Method for Purification of Human Hematopoietic Stem Cells by Flow Cytometry

Human hematopoietic stem cells (HSCs) and progenitors can be isolated by enriching for a rare cell population with a combination of monoclonal antibodies (MAbs). Such an isolation scheme involves multi-step procedures including ficoll-density fractionation and presort enric ...

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Isolation and Characterization of Primitive Hematopoietic Cells Based on Their Position in the Cell Cycle

Hematopoietic progenitor and stem cells are believed to lie dormant within the adult bone marrow microenvironment in a state characterized by both mitotic and metabolic quiescence. This state of cell-cycle quiescence has been the focus or target of many studies aimed at identifying cells ...

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Hematopoietic Colony-Forming Cells

The hematopoietic progenitors that can be assayed in clonal culture systems represent a continuum of differentiation, which includes multipotential progenitors and very late-committed progenitors with only limited cell-division capabilities (1). The late-committed pr ...

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Long-Term Culture-Initiating Cell Assays for Human and Murine Cells

In normal adults, the majority of primitive hematopoietic cells are concentrated in the bone marrow, where they are in contact with a variety of molecules that influence their cell-cycle status, viability, motility, and differentiation. These include components of the extracellular ...

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The Cobblestone-Area-Forming Cell Assay

Hematopoietic stem cell (HSC) subsets are defined by the capacity to which their offspring can contribute to the various mature blood-cell lineages. However, the proliferative potential of stem cells is highly dependent on the environment in which they reside, and it is only in retrospect th ...

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Isolation of Stromal Stem Cells From Human Adipose Tissue

The stromal compartment of mesenchymal tissues is thought to harbor stem cells that display extensive proliferative capacity and multilineage potential. Stromal stem cells offer a potentially large therapeutic potential in the field of regenerative medicine. Adipose tissue c ...

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Protocols for Nuclear Transfer in Mice

Cloning by nuclear transfer in mammals has revealed the remarkable ability of an oocyte to reprogram transferred cell nuclei and induce them to recapitulate the developmental program. This chapter summarizes the method used since 1998 for mouse cloning, which differs from that for large a ...

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RNAi-Based Methods for Gene Silencing in Mouse Oocytes

RNA interference (RNAi) is an evolutionary conserved gene-silencing pathway that can be efficiently utilized as a tool to study gene function. RNAi is initiated by long double-stranded RNAs (dsRNAs), which are processed into small duplexes called small-interfering RNAs (siRNAs). In t ...

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Making cRNA for Microinjection and Expression of Fluorescently Tagged Proteins for Live-Cell Imaging in Oocytes

Fluorescently tagged proteins have become a crucial weapon in the armory of a successful cell biology laboratory. This chapter describes how to produce cRNA coding for a fluorescently tagged protein of choice, such that it is suitable for microinjection and subsequent expression studi ...

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Using FRET to Study RanGTP Gradients in Live Mouse Oocytes

Oocytes are extremely large cells that have to coordinate accurate chromosome segregation, asymmetric cytoplasm partitioning together with their own development as fertilizable gametes. For this, they undergo both global (cell cycle progression related) and local changes. It ...

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Time-Lapse Epifluorescence Imaging of Expressed cRNA to Cyclin B1 for Studying Meiosis I in Mouse Oocytes

The first meiotic division of mammalian oocytes physiologically occurs in the ovary in the hours preceding ovulation. Fortunately, oocytes removed from their follicular environment will readily undergo this process in culture. Their large size, optical transparency, and effic ...

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